Cotton leaf curl virus resistance transgenics with antisense coat protein gene (AV1)

摘要

Cotton leaf curl virus disease is one of the major threats for cotton production and has emerged as a serious disease of cotton in North India. Cotton leaf curl disease (CLCuD) is caused by a geminivirus, transmitted by whitefly (Bemisia tabaci) vector. Genetic engineering of cotton transgenics resistant to CLCuD through antisense RNA approach is a potential technique to tackle the disease in cotton. Cotton transgenics resistant to CLCuD using antisense coat protein (ACP) gene was developed via Agrobacterium-mediated transformation. A binary vector carrying the ACP gene along with the nptII (neomycin phospho transferase) gene driven by CaMV-35S promoter and nos (nopaline synthase) terminator was used for transformation. Molecular confirmations of the transformants were carried by PCR and Southern analysis. The individual transgenics were raised in the greenhouse and screened for virus resistance by inoculating with viruliferous whiteflies. Following the challenge with the viruliferous whiteflies, transgenic plants remained symptomless. The T-2 plants were screened using specific primers for ACP gene and primers for nptII gene; they showed classical Mendelian pattern of inheritance.